Changes in the T and B lymphocyte subset profiles upon treatment of patients with Graves’ disease with radioactive iodine

The aim of the present study was to evaluate the subpopulation profile of T and B lymphocytes, and their relationships during therapy of the patients with Graves’ disease (GD) treated by means of radioactive iodine. We have examined 36 women with verified diagnosis of GD. The contents of thyroid hormones were determined by immunoradiometric analysis. The levels of thyroid-stimulating hormone receptor autoantibodies (rTSH) were evaluated by enzyme-linked immunosorbent assay. On the basis of comprehensive pre-therapeutic examination, all patients were exposed to the fixed-activity therapy with radioactive iodine-131 at a dose of 400 to 700 MBq administered orally in isotonic aqueous solution of sodium iodide. 56 practically healthy women were examined as a control group. The phenotype of T and B cells in whole blood was studied by flow cytometry using direct immunofluorescence. It was shown that the patients, prior to treatment with radioactive iodine, had high levels of cellular functional activity, as determined by expression of CD25 antigen on T cells and CD23-antigen on B lymphocytes. Higher functional activity of the cells responsive for adaptive immunity in the patients with GD manifests in the presence of increased levels of autoantibodies to rTSH. By means of correlation analysis, we found that the patients with GD examined before the therapy had the thyroid status may determine the functional stimulation of T and B cells, thus increasing the levels of autoimmune processes. One month after radioiodine therapy (RIT), the GD patients, along with transient hyperthyroidism with increased concentration of autoantibodies to rTSH, showed a reduction of activated T lymphocyte contents (including T helpers and cytotoxic T cells) to control values. However, the level of cytotoxic T lymphocytes in the blood remained low, and the content of Treg cells was significantly increased in the patients. Decreased contents of B cells activated memory B cell to the control levels were found in patients with GD over 1 month after RIT when studying the phenotype of blood B lymphocytes. In this case, increased levels of naive B lymphocytes and B2 cells were detected, as well as decreased numbers of activated B1 lymphocytes. The observed changes in the subpopulation composition of T and B cells, and in their phenotype developed against the background of complete absence of relationships between the studied parameters, thus suggesting loss of thyroid control of immune processes and cooperative cell interaction during the development of the immune response. Generally, the phenotypic changes of T and B lymphocyte subsets in the blood of patients with GD through 1 month after treatment with radioactive iodine may reflect a trend for decreased functional activity of adaptive cellular immunity which may also account for inhibition of autoimmune processes

REGULATORY INFLUENCE OF BLOOD MONOCYTES ON THE POPULATION COMPOSITION OF GRANULOCYTES AND THE STATE OF THEIR RESPIRATORY BURST IN THE WIDESPREAD PURULENT PERITONITIS

The aim of the study was to investigate the regulatory effect of monocytes and their subpopulations on the population composition  of granulocyte leukocytes and the state of their respiratory burst in  widespread purulent peritonitis (WPP). The study involved 24  patients aged 30-65 with acute surgical diseases and injuries of  abdominal organs complicated by WPP. As a control 25 relatively  healthy people of the same age range were examined. A study of the population composition of monocytes and granulocyte leukocytes in  blood was performed using a two-platform technology on the  hematological analyzer Sysmex XE-5000 (Sysmex Inc., USA) and  FC-500 flow cytometer (Beckman Coulter, USA) using the Cytodiff antibody kit (Beckman Coulter, USA). A study of the  monocytes number expressing HLA-DR- and CD64-receptor was performed by flow cytometry using direct immunofluorescence  of whole peripheral blood. The respiratory burst state of neutrophilic granulocytes was studied by chemiluminescence analysis on a 36-channel chemiluminescence analyzer BLM-3607 (MedBioTech, Russia). As indicators of chemiluminescence  were used luminol and lucigenin. The enhancement of  chemiluminescence induced by zymosan was evaluated by the ratio  of the area of the induced chemiluminescence to the spontaneous area and was defined as the activation index. It has  been established that the immune-inflammatory process in WPP is  characterized by a decrease in the number of classical monocytes in  the peripheral blood and an increase in the content of non-classical  monocytes. In WPP in peripheral blood the level of monocytes  expressing HLA-DR receptors decreases. The change in the ratio of  monocytes subpopulations characterizes the increase in the role of  the proinflammatory fraction in the WPP pathogenesis. Changes in  the population composition of granulocytes in the blood in patients  with WPP also characterize the development of an acute  inflammatory process. In this case, there is a decrease in the  number of basophils in the peripheral blood, which, apparently, is  determined by the presence of an allergic component in WPP and,  accordingly, their migration to the inflammation area. In patients  with WPP activation of a respiratory burst of granulocytes of blood  was detected, the intensity of which is determined by the synthesis  of primary and secondary active oxygen species. The results of the  correlation analysis made it possible to establish that in WPP the  regulatory role of non-classical monocytes increases aimed at  stimulating the inflammatory processes (an increase in the number  of mature and immature forms of neutrophils and stimulation of the activity of a respiratory explosion of granulocytes). The revealed features of the regulatory effect of monocytes on the population  composition and the intensity of the respiratory burst of granulocytes can be used in the development of immunotherapeutic methods aimed at reducing the activity of the inflammatory process in WPP

Recombinant Human Interleukin-2 Corrects NK Cell Phenotype and Functional Activity in Patients with Post-COVID Syndrome

Post-COVID syndrome develops in 10–20% of people who have recovered from COVID-19 and it is characterized by impaired function of the nervous, cardiovascular, and immune systems. Previously, it was found that patients who recovered from infection with the SARS-CoV-2 virus had a decrease in the number and functional activity of NK cells. The aim of the study was to assess the effectiveness of recombinant human IL-2 (rhIL-2) administered to correct NK cell phenotype and functional activity in patients with post-COVID syndrome. Patients were examined after 3 months for acute COVID-19 of varying severity. The phenotype of the peripheral blood NK cells was studied by flow cytometry. It was found that disturbances in the cell subset composition in patients with post-COVID syndrome were characterized by low levels of mature (p = 0.001) and cytotoxic NK cells (p = 0.013), with increased release of immature NK cells (p = 0.023). Functional deficiency of NK cells in post-COVID syndrome was characterized by lowered cytotoxic activity due to the decreased count of CD57+ (p = 0.001) and CD8+ (p < 0.001) NK cells. In the treatment of patients with post-COVID syndrome with recombinant IL-2, peripheral blood NK cell count and functional potential were restored. In general, the effectiveness of using rhIL-2 in treatment of post-COVID syndrome has been proven in patients with low levels of NK cells

BLOOD LEUKOCYTES PHENOTYPING BY HEMATOFLOW METHOD IN PATIENTS WITH ONYCHOMYCOSIS

The aim of the investigation was to evaluate the information content of Hematoflow method in the pathogenetic significance in determining the violations of the cellular responses of innate and adaptive immunity, as well as the decision on the appointment of immunotropic treatment of patients with onychomycosis. The study involved 42 patients with onychomycosis feet and hands/feet at the age of 20–45 years before the appointment of a systemic antifungal therapy. The diagnosis of mycosis, onychomycosis was confirmed by microscopic examination of the fragments of the damaged nail plate. The growth of the fungus culture on special media was observed in 64% of patients. 24 healthy persons were examined as controls. A study of the phenotype of white blood cells was performed on a dual-platform technology hematology analyzer and flow cytometry using a set of antibodies Cytodiff: CD36-FITC, CD2-PE, CD294(CRTH2)-PE, CD19-ECD, CD16-PC5 и CD45-PC7. The phenotypic composition evaluation of the white blood cells by the Hematoflow method allowed to establish in patients with onychomycosis the violation cellular innate and adaptive immunity. Minor changes were detected in the population composition of granule cells in the peripheral blood of patients manifested to an increase in the content of the young and segmented granulocytes. When monotsitopeniya patients with onychomycosis increases the content of the «classic» monocytes and decreases the level of «non-classical» monocytes. Changes in the composition of blood monocytes subpopulation identified in patients with the infection lasting up to 3 years and stored in the course of the disease. The most pronounced changes were found in patients with onychomycosis by the performance of adaptive immunity. Lymphopenia in these patients is realized by reducing the number of immature and mature B-cell, but by increasing the content of T-lymphocytes. Moreover, if the content of immature B-cells have decreased in patients with a duration of infection of up to 3 years, the change in the number of mature Tand B-lymphocytes detected during disease duration 3–10 and 10 years. These changes in the content of Tand B-lymphocytes reflect immunopathogenetic processes and determine the importance of Tand B-cell immunity in onychomycosis. In general, the Hematoflow method is informative in assessing violations the cell of innate and adaptive immunity. It allows to evaluate the severity of the immunopathological process mechanism and the level of damage to the immune system, can recommend its use for a personalized approach to the appointment immunotropic treatment